Table of contentsThe membrane stabilization method of human red blood cells (HRBC)Inhibition of protein denaturationAntiproteinase actionAnti-lipoxygenase activityThe membrane stabilization method of human red blood cells (HRBC)The blood was taken from a healthy human volunteer and An equal volume of Alsever's solution (2% dextrose, 0.8% sodium citrate, 0.5% citric acid and 0.42% NaCl) was was mixed and centrifuged at 3000 rpm for 10 min. The resulting concentrated cells were washed with normal saline and a 10% HRBC suspension was prepared. Different concentrations of AOAgNPs were prepared (50, 100, 150, 200 and 500 µg ml-1) using distilled water. A mixture of 1 ml phosphate buffer, 2 ml hypo saline and 0.5 ml HRBC suspension (at various concentrations mentioned above) was prepared. It was incubated for 30 minutes at 37°C and centrifuged at 3000 rpm for 20 minutes. The absorbance of the supernatant solution was measured spectrophotometrically at 560 nm (Kamalutheen et al., 2009). Diclofenac sodium was taken as standard medication. The experiment was repeated three times. Say no to plagiarism. Get a tailor-made essay on “Why violent video games should not be banned”?Get the original essayInhibition of protein denaturationTo study the inhibitory effect on protein denaturation, AOAgNPs were added to an aqueous solution at 1% BSA and the pH of the reaction mixture was adjusted. Sample mixtures were incubated at 37°C for 20 min and then heated again to 51°C for 20 min. After cooling, the turbidity of the samples was measured spectrophotometrically at 660 nm (Deshpande et al., 2009). Diclofenac sodium was used as standard medication. The experiment was carried out in triplicate. Antiproteinase action The antiproteinase activity of AOAgNPs was estimated according to the modified method of Sakat et al. 2010. Firstly, 2 ml of reaction mixture was prepared containing 0.06 mg/ml trypsin, 1 ml of 20 mM Tris HCl buffer (pH 7.4) and 1 ml of AOAgNP at different concentrations (100 - 500 µg ml-1). The mixture was incubated at 37°C for 5 min, then 1 ml of 0.8% (w/v) casein was added and incubated for 20 min. Then, to stop the reaction, 2 ml of 70% perchloric acid were added. Then, the entire suspension was centrifuged and the absorbance was measured at 210 nm. Indomethacin was used as the standard drug. The percentage inhibition of proteinase inhibitory activity was calculated using the following formula. Keep in mind: this is just a sample. Get a personalized article from our expert writers now. Get a Custom Assay Anti-lipoxygenase Activity The anti-lipoxygenase activity of AOAgNPs was studied using linoleic acid as substrate and lipoxidase as enzyme (Shinde et al. 1999). Samples were dissolved in 250 μl of 2 M borate buffer (pH 9.0) and 250 μl of lipoxidase enzyme solution (20,000 U/ml) and were incubated for 5 min at 25°C. After that, 1.0 mL of lenoleic acid solution (0.6 mM) was added, mixed well, and the absorbance was measured at 234 nm. Indomethacin was used as the standard drug and the inhibition percentage was calculated from the formula mentioned above..